人类滤泡树突状细胞的分离

Isolation of human follicular dendritic cell

目的 :为探讨人类滤泡树突细胞 (FDC)在HIV致病机制中的作用 ,摸索分离出提取人FDC的最佳方法。方法 :用胶原酶Ⅳ和DNA酶消化人扁桃体 ,Percoll连续梯度分层液分离含有FDC的细胞层 ,经MACS分选 ,得到富含FDC的细胞 ,用γ 射线将混有的B细胞、T细胞照射致死。结果 :流氏细胞仪分析FDC纯度约为 70 7% ,3H掺入法实验证明分离提取的细胞具有FDC细胞的活性 ,IL 6和IL 2的测定结果显示分离提取的细胞具有FDC的特征。结论 :此方法为一种行之有效的FDC分离提取方法 。

Objective:In order to study the role of FDC in HIV pathogenesis, an optimal method for isolating FDC was studied Methods:Tonsiles were digested with collagenase IV and Dnase I , and cellular layer including FDC was obtained by Percoll Purified FDC was yielded by MACS sorting ,and contaminated B and T cells were irradiated by γ ray Results:The purity of FDC analyzed with flow cytometer is about 70 72%, the thymidine incorporation showed that FDC cells with activity were yielded and the results of IL 6 & IL 2 determination indicated that isolated cells were with FDC characteristics Conclusion:This method is a practical method for isolating FDC for further investigation of FDC in HIV and neoplastic pathogenesis